Lissauer Matthew E, Johnson Steven B, Siuzdak Gary, Bochicchio Grant, Whiteford Craig, Nussbaumer Bill, Moore Richard, Scalea Thomas M
R. Adams Cowley Shock Trauma Center, University of Maryland Medical Center, Baltimore 21201, USA.
J Trauma. 2007 May;62(5):1082-92; discussion 1092-4. doi: 10.1097/TA.0b013e31804d23e1.
Systemic inflammatory response syndrome (SIRS) represents a host response to various insults. Recent advances have demonstrated an interconnection between inflammation, complement, and coagulation. This experiment was designed to evaluate differences in plasma protein profiles between clinically identical patients: septic versus uninfected SIRS patients, prior to clinical diagnosis of infection.
Patients admitted to an intensive care unit of a major university, meeting two of four SIRS criteria were followed prospectively for development of sepsis. Plasma samples were collected daily and divided into two groups: a preseptic group that subsequently developed sepsis and a SIRS group that remained uninfected. Protein profiling was accomplished by three-dimensional liquid chromatography fractionation with electrospray ion trap mass spectrometry after immunodepletion of abundant proteins and a trypsin digest. Spectra peaks were identified using Agilent Technologies Spectrum Mill Workbench software. Relevance to biologic pathways was analyzed and statistical significance determined with DAVID 2.1 available at the National Institutes of Health.
A total of 134 unique proteins were significantly different between groups. Thirty-two of these (23.5%) mapped to the complement and coagulation cascade (KEGG), 10 (7.5%) mapped to classic complement pathway; 11 (8.2%) mapped to complement pathway, and 8 (6.0%) mapped to lectin binding complement pathway (Biocarta). These pathways were all significantly (p<0.0001) over-represented in sepsis patients compared to SIRS-only patients.
Using novel mass spectrometry methodology, we were able to demonstrate differential protein profiles in septic versus uninfected SIRS patients prior to clinical diagnosis of sepsis.
全身炎症反应综合征（SIRS）代表机体对各种损伤的反应。最近的研究进展表明炎症、补体和凝血之间存在相互联系。本实验旨在评估临床特征相同的患者（脓毒症患者与未感染的SIRS患者）在感染临床诊断之前血浆蛋白谱的差异。
入住某大型大学重症监护病房、符合四项SIRS标准中的两项的患者被前瞻性地追踪观察脓毒症的发生情况。每天采集血浆样本并分为两组：随后发生脓毒症的脓毒症前期组和未发生感染的SIRS组。在去除丰富蛋白质并进行胰蛋白酶消化后，通过三维液相色谱分离结合电喷雾离子阱质谱对蛋白质进行分析。使用安捷伦科技公司的Spectrum Mill Workbench软件识别光谱峰。利用美国国立卫生研究院提供的DAVID 2.1分析与生物途径的相关性并确定统计学意义。
两组之间共有134种独特的蛋白质存在显著差异。其中32种（23.5%）映射到补体和凝血级联反应（KEGG），10种（7.5%）映射到经典补体途径；11种（8.2%）映射到补体途径，8种（6.0%）映射到凝集素结合补体途径（Biocarta）。与仅患有SIRS的患者相比，这些途径在脓毒症患者中均显著（p < 0.0001）过度表达。
使用新型质谱方法，我们能够在脓毒症临床诊断之前证明脓毒症患者与未感染的SIRS患者之间存在不同的蛋白质谱。
